Sensitive to Apoptosis Gene (SAG, human; Sag, mouse) also known as RBX2/ROC2, is an essential RING component of SCF (SKP1, Cullins, F-box proteins) E3 ubiquitin ligase which, by promoting ubiquitination and degradation of various key regulatory proteins, controls several important biological processes including cell cycle progression, signal transduction, transcription, and DNA replication. Our published and unpublished data showed a direct association of SAG with pancreatic cancer as follows: a) SAG is overexpressed in pancreatic cancer tissues, and SAG nuclear expression is associated with poor prognosis of pancreatic cancer patients; b) SAG siRNA knockdown caused accumulation of tumor suppressive proteins and inhibited survival and anchorage-independent growth of pancreatic cancer cells, and c) orthotopic in vivo growth and matrigel invasion of pancreatic cancer cells were significantly inhibited, respectively, by SAG siRNA knockdown. All these studies suggested that SAG overexpression could be oncogenic during pancreatic tumorigenesis. The objectives of this application are to use pancreatic specific Sag transgenic or KO mouse models to mechanistically study the role of Sag in pancreatic tumorigenesis triggered by Kras activation (KrasG12D) and p53 mutation (p53R172H). The central hypothesis is that SAG promotes tumorigenesis by promoting the degradation of tumor suppressive substrates such as DEPTOR, IB, and p21/p27, leading to activation of the mTOR and NFB pathways. Thus, SAG-pancreatic transgenic expression would promote pancreatic tumorigenesis, initiated by KrasG12D, whereas Sag pancreatic deletion would abrogate oncogenic signals by accumulation of tumor suppressive proteins, thus suppressing pancreatic tumorigenesis triggered by KrasG12D and p53R172H. Two specific aims are proposed to 1) determine the role of Sag in KrasG12D/p53R172H-induced pancreatic tumorigenesis and 2) elucidate mechanism(s) by which Sag regulates pancreatic tumorigenesis. IMPACT: Our study uses mouse models that recapitulate the development of pancreatic ductal adenocarcinoma (PDAC) to investigate the role of Sag E3 ligase in the initiation and progression of PDAC and to elucidate its mechanism of action. Our study will mechanistically validate SAG E3 ubiquitin ligase as a novel anti-pancreatic cancer target and provide proof-of-concept evidence for future discovery of specific inhibitor of SAG E3 ligase as a novel class of anti-pancreatic cancer drugs. Our work is, therefore, highly innovative and of significant impact with translational value.